日本血吸虫Sj22.6-LHD-Sj23融合基因的克隆及原核表达  

Cloning and prokaryotic expression of the Schistosoma japonicum Sj22.6-LHD-Sj23 fusion gene

作　　者：付媛[1] 卢福庄[1] 石团员[1] 顾昀[1] 俞国乔 陆国林 张雪娟[1] 

机构地区：[1]浙江省农业科学院畜牧兽医研究所,浙江杭州310021 [2]浙江省畜牧兽医局,浙江杭州310020

出　　处：《中国预防兽医学报》2010年第11期904-907,共4页Chinese Journal of Preventive Veterinary Medicine

基　　金：浙江省攻关项目资助

摘　　要：为获得具有良好抗原性的日本血吸虫重组蛋白,本研究设计了两对特异性引物,以日本血吸虫mRNA为模板,RT-PCR方法分别克隆基因片段Sj22.6ku和LHD-Sj23,以重叠延伸PCR方法将两个片段连接,构建原核重组表达质粒pET28-Sj22.6-LHD-Sj23;该重组质粒在大肠杆菌Rosetta DE3中表达后所获得的融合蛋白Sj22.6-LHD-Sj23分子量约为35 ku。Western blot鉴定结果表明该重组蛋白有良好的免疫原性,为进一步研制日本血吸虫病分子诊断试剂盒奠定了基础。In this study,cDNA fragments of Sj22.6 ku and LHD-Sj23 gene of Schistosoma japonicum were amplified by specific primers,respectively,from mRNA of S.japonicum by RT-PCR,and fused together by overlap-extension PCR.The fusion gene was cloned into prokaryotic expression plasmid to construct pET28-Sj22.6-LHD-Sj23 and transformed to E.coli Rosetta DE3 cells.The recombinant protein approx 35 ku was expressed and its antigenicity was confirmed by western blot.

关 键 词：日本血吸虫 Sj22.6ku蛋白 LHD-Sj23蛋白 融合蛋白 重叠延伸PCR 

分 类 号：S852.7[农业科学—基础兽医学]